Method Description

Methemoglobin:

The normal absorption spectrum of oxyhemoglobin has very little optical density above 600 nm. The absorption spectrum of methemoglobin exhibits a small, characteristic peak at 630 nm. This peak is abolished as methemoglobin is converted to cyanmethemoglobin upon addition of potassium cyanide, and the drop in optical density is proportional to methemoglobin concentration.

Sulfhemoglobin:

The normal absorption spectrum of oxyhemoglobin has very little optical density above 600 nm. However, if certain poorly defined hemoglobin denaturation products are present in a hemolysate, there is a broad elevation of the absorption curve in the range of 600 nm to 620 nm. This sulfhemoglobin plateau is not affected by treatment with cyanide. Sulfhemoglobin is not available, nor can it be prepared, in a pure form for preparation of a sulfhemoglobin standard. In calculating sulfhemoglobin concentration, the factor for sulfhemoglobin quantitation is based on studies of Carrico, et al.(Evelyn KA, Malloy HT. Microdetermination of oxyhemoglobin, methemoglobin, and sulfhemoglobin in a single sample of blood. J Biol Chem. 1938;126:655-662; Carrico RJ, Peisach J, Alben JO. The preparation and some physical properties of sulfhemoglobin. J Analyt Biochem. 1978;253[10]:2386-2391; So JCC, Ma ESK. Hemoglobin and hemoglobinopathies. In: Rifai N, Chiu RWK, Young I, Burnham CAD, Wittwer CT, eds. Tietz Textbook of Laboratory Medicine. Elsevier; 2023:chap 77)